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The cells were then resuspended in cortical neuron culture medium (CM) consisting of Neurobasal medium (Gibco, 10888022) supplemented with B-27 supplement (Gibco, 17504044), 10 ng/mL NT-3 (PeproTech, 450-03), 10 ng/mL BDNF (PeproTech, 450-02), 1 µg/mL laminin (Sigma-Aldrich, L2020), 2 µg/mL doxycycline (Sigma, D9891), and 10 µM Y-27632 ROCK inhibitor and seeded to the PLO-coated surface